Surfactants exhibit critical functionalities such as emulsifiers, dispersants, and foaming agents due to their amphiphilic nature and capacity to modulate interfacial phenomena. While synthetic surfactants dominate industrial applications, their persistence and ecotoxicity underscore an urgent demand for biodegradable, low-toxicity alternatives. In this context, biosurfactants have emerged as multifunctional and environmentally benign substitutes. Microbial biosurfactants, including glycolipids and lipopeptides synthesized by diverse bacterial and fungal taxa, offer scalable production and structural versatility. These compounds demonstrate interesting surface properties, low critical micelle concentrations, and resilience under extreme environmental conditions, enabling broad utility across industrial sectors. Notably, microbial biosurfactants possess distinctive functionalities that extend beyond surface modulation. In agri-food systems, they improve rheological and organoleptic properties, exhibit antimicrobial and antioxidant activities, and contribute to sustainable intensification by enhancing soil fertility, plant health, and resistance to biotic stressors. Biosurfactants derived from Generally Recognized As safe (GRAS) microorganisms are especially promising for integration into food-grade and pharmaceutical formulations. Furthermore, their capacity to degrade organic pollutants and stimulate indigenous microbial communities positions them as strategic agents in environmental remediation. This chapter presents an integrative and up-to-date examination of microbial biosurfactants, emphasizing their structural diversity, biosynthetic potential, and applicability in sustainable agriculture and food systems. It critically assesses current research, market trajectories, and technological bottlenecks, thereby offering comprehensive insights into their role within a transitioning bioeconomy.